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Structure-function studies of nucleocytoplasmic transport of retroviral genomic RNA by mRNA export factor TAP

Nature Structural & Molecular Biology volume 18, pages 990998 (2011) | Download Citation

Abstract

mRNA export is mediated by the TAP–p15 heterodimer, which belongs to the family of NTF2-like export receptors. TAP–p15 heterodimers also bind to the constitutive transport element (CTE) present in simian type D retroviral RNAs, and they mediate the export of viral unspliced RNAs to the host cytoplasm. We have solved the crystal structure of the RNA recognition and leucine-rich repeat motifs of TAP bound to one symmetrical half of the CTE RNA. L-shaped conformations of protein and RNA are involved in a mutual molecular embrace on complex formation. We have monitored the impact of structure-guided mutations on binding affinities in vitro and transport assays in vivo. Our studies define the principles by which CTE RNA subverts the mRNA export receptor TAP, thereby facilitating the nuclear export of viral genomic RNAs, and, more generally, provide insights on cargo RNA recognition by mRNA export receptors.

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Acknowledgements

This research was supported by funds from the US National Institutes of Health (CA049982) to D.J.P. and the Max-Planck Society to E.I. X-ray data were collected at the X-29 beamline at the National Synchrotron Light Source at the Brookhaven National Laboratory and we are grateful to the staff for their assistance.

Author information

Affiliations

  1. Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.

    • Marianna Teplova
    • , Nyan W Khin
    •  & Dinshaw J Patel
  2. Max Planck Institute for Developmental Biology, Tübingen, Germany.

    • Lara Wohlbold
    •  & Elisa Izaurralde

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Contributions

Constructs design, protein and RNA preparation and purification, crystallization of complex and its structure determination and in vitro binding assays were undertaken by M.T. with the assistance of N.W.K. under the supervision of D.J.P. The in vivo transport assays were performed by L.W. under the supervision of E.I. The paper was written by M.T., D.J.P. and E.I. with input from the other authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Dinshaw J Patel.

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https://doi.org/10.1038/nsmb.2094

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